KR20130095244A - Phosphaplatins and their use for treatment of cancers - Google Patents

Abstract

Stable monomeric phosphaplatin, ie (pyrophosphato) platinum (II) or platinum (IV), comprising a cis - cyclohexanediamine ligand or an enantiomerically enriched or enantiomerically pure trans- cyclohexanediamine ligand. ) Complexes and methods of synthesizing these complexes are provided. The efficacy and toxicity of the phosphaplatin compounds are measured for various cancers, including sensitive and resistant ovarian cancer, head and neck cancer, and colon cancer. Compositions comprising the platinum complexes and methods of treating proliferative diseases or disorders with the complexes or compositions comprising the same are described.

Description

Phosphoplatin and its use to treat cancer {PHOSPHAPLATINS AND THEIR USE FOR TREATMENT OF CANCERS}

Cross Reference of Related Applications

This application is directed to 35 U.S.C. US patent application Ser. No. 61 / 351,514, filed June 4, 2010. Claim priority under §119 (e), which is incorporated herein by reference in its entirety.

Field of invention

The present application generally relates to pyrophosphato platinum complexes; A method of synthesizing the provided complexes; A composition comprising the complex provided above; And methods of treating proliferative diseases using the provided complexes, compositions comprising said provided complexes, or combinations thereof.

In 2008, more than 12,000,000 people were diagnosed with cancer and more than 7,000,000 people died of cancer worldwide. In fact, cancer is the leading cause of death in developed countries and the second leading cause of death in developing countries (second only for HIV / AIDS). Once diagnosed with cancer, the patient's prognosis depends largely on factors such as whether the cancer has been diagnosed at an early stage, whether the cancer has spread throughout the body, and whether the cancer has become or has become resistant to known chemotherapy regimens.

Platinum-based anticancer drugs cisplatin, carboplatin and oxaliplatin are widely used to treat various cancers such as ovarian cancer, testicular cancer, small cell lung cancer and colorectal cancer. These compounds can be used in combination with other treatment regimens, including radiation therapy, to treat a wide variety of cancers. Currently, more than 600 clinical trials in adjuvant treatment modalities using platinum compounds highlight the potential of platinum compounds in effectively treating a wide variety of other cancers. For example, recent breakthroughs suggest that the diabetic drug rosiglitazone can be effectively used in combination with carboplatin to treat various forms of cancer. Currently, this presentation adds a new perspective to the ever-increasing application of platinum-based anticancer drugs because most adjuvant therapies are primarily limited to the combination of cancer or radiation drugs with other cancer drugs. Thus, there is a continuing need for new platinum-based anticancer drugs as well as new applications for platinum-based anticancer drugs.

Conventional platinum chemotherapeutic agents such as cisplatin initiate apoptosis mainly at the G2 stage of the cell cycle, mainly through transcriptional inhibition and replication inhibition processes, especially at high concentrations. Covalent binding of guanine and adenine bases to the DNA through the N7 site by both intra-strand and inter-strand modes leads to a cascade of cellular responses leading to apoptosis (programmed cell death). It is thought to be a major molecular event. Numerous problems have been identified in understanding the complexity of the molecular mechanisms of cisplatin's cellular and molecular metal-biochemistry and cytotoxicity. Briefly, it was noted that platinumed DNA is at the heart of the onset of cytotoxicity. Platinum-bound DNA is isolated from repair by nucleotide excision repair (NER) enzyme by high mobility protein (HMG). In addition, such platinum-DNA adducts are believed to activate p53 transcription factors, induce histone phosphorylation and induce chromatin condensation.

Although platinum-based chemotherapeutic agents are widely used to treat cancer, their application is limited in many patients because of their nephrotoxicity, neurotoxicity, endotoxicity, myelosuppression and acquired resistance to platinum-metal drugs. For example, a significant proportion of patients become resistant to cisplatin treatment. Carboplatin somewhat reduces toxicity compared to cisplatin, but does not alleviate resistance. Recently, oxaliplatin has been recognized to treat colorectal cancer, but its resistance has not been investigated much.

The art lacks an understanding of the molecular development of resistance to conventional platinum-based chemotherapeutic agents and methods for overcoming such resistance. Although not completely understood, the ability to repair DNA damage by cleaving bound platinum from DNA is thought to contribute to resistance mechanisms. Other mechanisms involved in contributing to resistance include reduced intracellular accumulation of cisplatin due to reduced uptake associated with down-regulation of expression of the copper transport protein CTR1; increased outflow due to overexpression of cMOAT, ATP7A and ATP7B; Impaired down regulation of apoptosis pro-apoptotic genes; And up-regulation of anti-apoptotic genes. On the other hand, up-regulation of CTR1 protein is associated with increased endotoxin. In addition, replacement of MAPK and inactivation of platinum by glutathione and other small molecules and proteins, in particular metallothionine, have been assumed to render the factors resistant to platinum drugs. In view of the above, there is a need for a method for overcoming resistance to platinum drugs, including the development of drugs that are not susceptible to DNA repair mechanisms.

In various embodiments, U.S. Pat. Patent No. 7,700,649 (Bose) and U.S. Serial No. 12 / 722,189 (Bose) addresses the needs of the art by describing synthetic routes and methods of treating cancer comprising a new class of platinum complexes, namely pyrophosphato complexes with platinum (II) or platinum (IV) metal centers. Meets some. The compounds and methods described herein are part of a drug development strategy based on the manufacture of a class of platinum anti-tumor agents that negate DNA-recovery based resistance by not covalently binding to DNA. This strategy is a paradigm shift from the existing platinum drug development approach, which is a major theme in the development of platinum anticancer agents, where DNA binding is more effective.

Racemic trans- (±) -1,2 - cyclohexanediamine (pyrophosphato) platinum (II) and racemic trans- (±) -1,2 - cyclohexanediamine- trans -dihydroxy (pyrophosphato) ) Platinum (IV) is described in US Patent No. 7,700,649 (Bose) and US Serial No. 12 / 722,189 (Bose), among the pyrophosphato platinum complexes. Although these racemic complexes have been found to be effective for the treatment of some cancers, cancer cell growth by improving the efficacy of pyrophosphato platinum therapeutics, reducing the toxicity of these therapeutics, and improving targeting of genes involved in killing cancer cells. There is still a need for an improved drug development approach, including but not limited to inhibiting these.

In various embodiments, the present disclosure addresses drug development strategies by describing drug development strategies based on enantiomeric pure monomeric pyrophosphato platinum complexes and enantioenriched monomeric pyrophosphato platinum complexes. do. Thus, the present technology not only identifies target genes involved in killing cancer cells and inhibits cancer cell growth, but also provides the most effective form of pyrophosphato platinum complexes. The complexes provided are stable, exhibit improved cytotoxicity and are more effective than conventional anticancer agents. In addition, this drug development strategy is a paradigm shift from the conventional platinum drug development approach where DNA binding remains the main theme.

In various embodiments, the present invention provides isolated monomeric (( cis or trans ) -1,2-cyclohexanediamine) (dihydrogen pyrophosphato) platinum (II) and (( cis or trans ) -1,2 -Cyclohexanediamine) -trans -dihydroxy (dihydrogen pyrophosphato) platinum (IV) complex, wherein the complex is enantiomerically pure or enantiomeric excess of cis- 1 Or a 2-cyclohexanediamine-based complex or includes one of two distinguishable trans -1,2-cyclohexanediamine-based complexes. Thus, the present technology is based on (i) ((1 R , 2 R ) -1,2-cyclohexanediamine) (dihydrogen pyrophosphato) platinum (II) (herein "(1 R , 2 R ) -fatigue Referred to as dark-2 "; in "fatigue dark -2 (1 S, 2 S) " (ii) ((1 S, 2 S) -1,2- cyclohexane diamine) (dihydrogen fatigue force pato) platinum (II) (present in Mentioned); (iii) ((1 R, 2 S) -1,2- cyclohexane diamine) (dihydrogen fatigue force pato) platinum (II) or ((1 S, 2 R) -1,2- cyclohexane-diamine) (Dihydrogen pyrophosphato) platinum (II) (overlapping enantiomeric compound and collectively referred to herein as “ cis -pyrodak-2”); (iv) ((1 R, 2 R) -1,2- cyclohexane-diamine) -trans-dihydroxy rokso (dihydrogen fatigue force pato) platinum (IV) ( "(1 R , 2 R herein) Referred to as pyrodark-4 "; (v) ((1 S, 2 S) -1,2- cyclohexane-diamine) -trans-dihydroxy rokso (dihydrogen fatigue force pato) platinum (IV) (herein "(1 S, 2 S) - Referred to as pyrodark-4 "; And (vi) ((1 R, 2 S) -1,2- cyclohexane-diamine) -trans-dihydroxy rokso (dihydrogen fatigue force pato) platinum (IV) or ((1 S, 2 R) -1 , 2-cyclohexanediamine) -trans -dihydroxy (dihydrogen pyrophosphato) platinum (IV) (overlapping enantiomeric compound and collectively referred to herein as " cis -pyrodak-4"); And platinum (II) and platinum (IV) complexes selected from pharmaceutically acceptable salts or solvates of any one of (i)-(vi). With respect to the amino groups on the 1,2-cyclohexanediamine ligands in compounds (i), (ii), (iv) and (v), the (1 R , 2 R ) and (1 S , 2 S ) stereochemistry Amino groups of the trans configuration are shown, and in compounds (iii) and (vi) the (1 R , 2 S ) and (1 S , 2 R ) stereochemistry represents the amino group of the cis configuration.

In addition, the present invention provides compositions in some embodiments comprising a therapeutically effective amount of one or more provided complexes and one or more pharmaceutically acceptable ingredients, such as a carrier, diluent, adjuvant or vehicle.

In another embodiment, the present technology provides a method of treating one or more proliferative diseases by administering a therapeutically effective amount of a composition comprising one or more of the complexes provided in a subject in need thereof.

Although the present specification concludes with the claims that particularly point out and specifically claim the invention, it is believed that the invention will be better understood from the following description in conjunction with the accompanying drawings.
1 shows (I) (1 R , 2 R ) -pyrodak-2, (II) (1 S , 2 S ) -pyrodak-2, (III) cis -pyrodak-2, (IV) (1 R, 2 R) - dark fatigue -4, (V) (1 S , 2 S) - fatigue dark -4, (VI) cis-fatigue dark-4, and the trans - (±) - -2 of fatigue dark solid Circular circular dichroism spectrum of the isomer is shown.
FIG. 2 shows (1 R , 2 R ) -pyrodak-2, (1 S , 2 S ) -pyrodak measured from a clonality assay by exposing human ovarian cancer cells (A2780) to various concentrations of compounds for 24 hours. The activity of -2 and trans- (±) -pyrodak-2 is shown.
FIG. 3 shows (1 R , 2 R ) -pyrrodak-2, (1 S , measured from a clonality assay by exposing cisplatin-resistant human ovarian cancer cells (OVCAR-10) to various concentrations of compounds for 24 hours. 2 S ) -Pyrrodak-2 and trans- (±) -pyrodak-2 are shown.
4 is a plot of mean tumor size over a six week period during phosphaplatin administration according to an embodiment described herein for human ovarian cancer cells in mice.
Blood dark-4 - Figure 5 is cisplatin-resistant human ovarian cancer cells, are described in detail in the following, for the (OVCAR-10) (1 R , 2 R) - fatigue dark-2 and (1 R, 2 R) It shows efficiency.
FIG. 6 is a control (PBS / Bic) administered once every other day (“qodx3”) for 3 days, carboplatin administered at 60 mg / kg (“qodx3” administration), and 40 mg / kg [3 days A plot of tumor size over time comparing successive once-daily administrations (“qdx”)] (1 R , 2 R ) -Pyridak-4 is shown.
FIG. 7 shows (1 R , 2 R ) -pyrodak-2 [administered qodx3 and once daily for four consecutive days (“qdx4”)) for human head and neck cancer (UMSCC10b) and The efficiency of (1 R , 2 R ) -pyrrodac-4 (administered qdx4) is shown.

Specific embodiments of the present technology will be described. However, the invention may be embodied in different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this technology will be thorough and will fully convey the scope of the invention to those skilled in the art.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for describing particular embodiments only and is not intended to be limiting of the invention. As used in this specification and claims, the singular forms "a", "an" and "the" are intended to include the plural forms unless the context clearly dictates otherwise.

Terms such as "preferably", "generally" and "typically" are used to limit the scope of the claimed invention or to mean that certain features are critical, essential or even important for the structure or function of the claimed invention. It should be noted that not. Rather, these terms are only intended to highlight alternative or additional features that may or may not be used in certain embodiments of the present invention.

The term "substantially" is used herein to denote inherent uncertainty that may be due to any quantitative comparison, value, measurement or other expression. Alternatively, the term “substantially” is used herein to indicate that quantitative expression can change from a defined criterion without causing a change in the underlying function of the subject matter in question. As such, the term is expected to show an exact relationship or aspect in theory, but in practice any quantitative comparison, value, measurement or other, referring to an element or feature of a batch that may embody something slightly less accurately. Used to indicate inherent incompatibilities that may be due to presentation.

Unless otherwise indicated, all numbers expressing quantities of ingredients, properties such as molecular weight, reaction conditions, etc., as used in this specification and claims are to be understood as being modified in all instances by the term "about", It is intended to mean up to ± 10% of the indicated values. In addition, it is to be understood that the description of any range in this specification and claims includes not only the endpoint but also the range itself and those encompassed within the scope. Unless otherwise indicated, the numerical characteristics set forth herein and in the claims are approximations that may vary depending on the desired characteristics to be obtained in the embodiments of the present invention. Notwithstanding that the numerical ranges and variables setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. However, any numerical value inherently retains certain errors resulting from the errors found in each measurement.

As used herein, the term “phosphaplatin” generally refers to a platinum complex that is coordinated with a single bidentate pyrophosphato ligand. Phosphoplatin according to embodiments described herein may have the general structural formulas of Formulas (A) and (B):

(A)

[Formula B]

Wherein L ¹ and L ² are independently selected from a neutral ligand (NH ₃ ; substituted or unsubstituted aliphatic amine; and substituted or unsubstituted aromatic amine) coordinated to the platinum metal center, or terminal group L ¹ And a single bidentate neutral ligand having L ² (selected from substituted or unsubstituted aliphatic or aromatic diamines); L ³ and L ⁴ are ligands coordinated to the platinum metal center (selected from hydroxides, acetic acid, butyric acid, and alpha-hydroxy acids, amines or charged species thereof). Pyrophosphato ligands may be neutral (not shown) or charged (shown). When charged, pyrophosphato ligands are present with counter ions represented by Z ⁺ . Examples of Z ⁺ include hydrogen; Alkali metals such as sodium and potassium; And monovalent organic residues. Preferably, Z ⁺ is a counter ion resulting in a pharmaceutically acceptable salt. Regardless of whether it is charged or neutral, the general structure of the platinum (II) complex represented by formula (A) is square-plane, and the general structure of the platinum (IV) complex represented by formula (B) is octahedron.

In general, phosphaplatin is not readily hydrolyzed, soluble in aqueous solutions of neutral pH and stable in aqueous solutions of neutral pH. In addition, phosphaplatin is generally cytotoxic in cancer cell lines and is effective in cell lines that are resistant to one or both of cisplatin and carboplatin. Thus, phosphaplatin is effective and in some cases more effective in inducing cancer cell death compared to known platinum cancer drugs and exhibits the desired stability and solubility in solution, suitable for administration to a patient. As used herein in connection with the phosphaplatin of the present invention, the term "stable" refers to hydrolysis resistance when the complex is maintained in an aqueous solution of pH 6-8 for a period of 2-6 days.

Unlike cisplatin, carboplatin, and related platinum based anticancer agents, phosphaplatin does not covalently bind DNA. Resistance to cisplatin, carboplatin, and related platinum anticancer agents is believed to result from effective repair of DNA damage by various enzymes, including nuclear ablation repair enzymes. However, since phosphaplatin does not covalently bind DNA, resistance to phosphaplatin due to DNA repair mechanisms will be impossible. The data suggest that phosphaplatin causes overexpression of fas and fas-related transcription factors, some apoptosis promoting genes such as Bak and Bax, and tumor suppressor genes such as PUMA and PTEN. Phosphoplatin also down-regulates BCL2, an antiapoptotic gene. Western blot experiments dealing with the expression of proteins transcribed by these genes show parallel trends. In addition, although the cell binding of phosphaplatin is less than cisplatin, phosphaplatin shows high cytotoxicity. Accordingly, the present invention provides effective platinum anticancer agents with molecular targets different from those in the art.

The term "enatiomeric excess" is used herein according to its generally understood definition. That is, for two enantiomers A and B, which may be present in the mixture in molar amounts of M _A and M _B , respectively, the enantiomeric excess E for the enantiomer present in the higher molar amount in the mixture is represented by the formula% E. = | (M _A -M _B ) / (M _A + M _B ) × 100 |, where E is greater than 0%. The "racemic mixture" of enantiomers A and B, which may be represented by abbreviations such as " rac " and / or "(±)" (or simply without any reference to enantiomers), is M _A = M _B Because of this, it has an E of 0%. As described further herein, a mixture consisting of the A and B M _A and M _B is 60%, 40% has an enantiomerically an excess of A is equivalent to 20%. In the same mixture, each molecule of B (40% of the mixture) is paired with the A molecule (40% of the mixture) in the mixture, so that excess A molecules (20% of the mixture) can be left unpaired. It can be considered as a mixture consisting of 80% racemic mixtures of A and B with 20% of thiomerically pure A.

The term "enantiomeric" as used herein for a molecule having two enantiomers A and B includes substantially only one of enantiomers A or B but not both A and B. Does not refer to a compound or composition. For an “enantiomerically pure” complex, 97% ≦ E ≦ 100%.

As used herein, the term “enantioenriched” means, in a broad sense, that a compound or composition comprising a molecule having two enantiomers A and B is referred to as enantiomeric A or B. Refers to a compound or composition having a cumulative excess. Thus, an "enantiomeric mixture of A and B" refers to a mixture having 0% <E ≤ 100% for A or B and having an enantiomeric excess of A or a mixture with enantiomeric excess of B. Can be. As an illustrative example, the enantiomeric excess of A or B is greater than 0.01%, greater than 1%, greater than 10%, greater than 25%, greater than 50%, greater than 75%, greater than 90%, greater than 98%, greater than 99%, It may be greater than 99.9% or even 100%.

In various embodiments, provided herein are stable monomeric phosphaplatin complexes (and compositions comprising a therapeutically effective amount of one or more such complexes). In some embodiments, the complexes and compositions can be used in methods of treating cancers including but not limited to cancers resistant to treatment with one or more of cisplatin, carboplatin and oxaliplatin.

Complex

In various embodiments, phosphaplatin complexes are selected from the group consisting of:

(i) enantiomerically pure (1 R , 2 R ) -pyrodak-2 of Formula I:

(I)

(I);

(I);

(ii) Pure enantiomerically (1 S, 2 S) of formula II - fatigue dark-2:

&Lt; RTI ID = 0.0 &

(II);

(II);

(iii) enantiomerically rich pyridac-2 having an enantiomeric excess of (1 R , 2 R ) -pyrodak-2 of formula (I) or (1 S , 2 S ) -pyrodak-2 of formula (II) :

(I)

&Lt; RTI ID = 0.0 &

;

;

(iv) cis -pyrodak-2 of formula III:

[Formula III]

;

;

(v) enantiomerically pure (1 R , 2 R ) -pyrodak-4 of Formula IV:

(IV)

;

;

(vi) Pure enantiomerically of formula V (1 S, 2 S) - Dark fatigue 4:

(V)

;

;

(vii) Enantiomerically rich pyridac-4 having an enantiomeric excess of (1 R , 2 R ) -pyrodak-4 of formula IV or (1 S , 2 S ) -pyrodak-4 of formula V :

(IV)

(V)

; 및

; And

(viii) cis -pyrodak-4 of formula VI:

(VI)

.

.

In the complexes according to formulas (I) to (VI), the abbreviation "pyrodach" refers to a 1,2-cyclohexanediamine (pyrophosphato) platinum complex, wherein "pyro" refers to a coordination pyrophosphato ligand, "Dark" refers to a coordination 1,2-cyclohexanediamine ligand (named according to the IUPAC Convention), also known as 1,2-diaminocyclohexane. The term (1 R , 2 R ), (1 S , 2 S ) or cis before the term "pyrrodak" is a stereochemical arrangement of chiral centers at the 1- and 2-positions of 1,2-cyclohexanediamine ligands Say The number (ie 2 or 4) after the indication "pyrodak" refers to the oxidation number of the platinum center. That is, "pyrodak-2" refers to a platinum (II) complex and "pyrodak-4" refers to a platinum (IV) complex.

Pyrophosphate and 1,2-cyclohexane general formulas (I) to force papeul Latin of formula VI with a diamine ligand is coordinated to platinum in the two amino chiral center at the carbon of the diamine ligand 1 and ₂ (-NH 2) group of possible cis- And trans -geometric structures may exist in four stereoisomers. These stereoisomers (1 R, 2 R) - , (1 S, 2 S) -, (1 R, 2 S) - and (1 S, 2 R) - shows the arrangement. The ligand trans-1,2-trans-cyclohexane diamine (1 R, 2 R) respectively to provide the two enantiomers having the arrangement - and (1 S, 2 S). Cis-isomer, in principle, (1 R, 2 S) - possible isomers are overlapped with each other structural and chemical that can not be separated, even - and (1 S, 2 R) - Ahira including thio Murray However, these two cis It is a mirror image. Thus, the cis-isomer since the two enantiomers of simply "cis-isomer" shall indicate in, is referred to with a single formula.

Enantiomerically rich pyridak-2 mixture (iii) and enantiomerically rich pyridak-4 mixture (vii) are both enantiomeric excess of (1 R , 2 R ) -enantiomer or (1 S, 2 S) - enantiomers are characterized meoro. Enantiomeric excess may vary, and in exemplary embodiments greater than 0.01%, greater than 1%, greater than 10%, greater than 25%, greater than 50%, greater than 75%, greater than 90%, greater than 98%, greater than 99% , Greater than 99.9% or even 100%. In an exemplary embodiment, the enantiomeric excess is an enantiomeric excess of (1 R , 2 R ) -enantiomer, for example greater than 90% of (1 R , 2 R ) -enantiomer. It is an overdose. In further exemplary embodiments, the enantiomeric excess is an enantiomeric excess of (1 S , 2 S ) -enantiomer, for example greater than 90% of the (1 S , 2 S ) -enantiomer. Enantiomeric excess. In a further exemplary embodiment, the enantiomerically enriched pyridak-2 mixture (i) and / or the enantiomerically enriched pyridak-4 mixture (ii) is a (1 R , 2 R ) -enantiomer or it is enantiomerically pure for the enantiomer - (1 S, 2 S) .

Enantiomerically pure complexes according to formulas (I), (II), (IV) and (V) presented herein are described in US Pat. No. 7,700,649, in the comparative data between the corresponding racemic mixtures described in, the racemic mixtures of (1 R , 2 R ) -pyrodak-2 and (1 S , 2 S ) -pyrodak-2 are abbreviated as " trans- ( ±) -pyrrodak-2. Likewise, a racemic mixture of (1 R , 2 R ) -pyrodak-4 and (1 S , 2 S ) -pyrodak-4 is abbreviated as " trans- (± Will be referred to as pyrodark-4.

By way of non-limiting example, compounds of formulas (I)-(VI) can be prepared by adding potassium iodide to convert K ₂ PtCl ₄ to K ₂ PtI ₄ and cis- (1,2-cyclohexanediamine) dichloroplatinum (II); It can be synthesized from the same starting material. Then, K ₂ PtI ₄ is cis-1,2-cyclohexane diamine, trans - (1 R, 2 R) -1, 2-cyclohexane diamine, trans - (1 S, 2 S) -1, 2-cyclopenten React with 1,2-cyclohexanediamine with preferred stereochemistry such as hexanediamine, or mixtures thereof. The resulting (1,2-cyclohexane-diamine) di-iodo-platinum (II) complexes by the addition of 2 equivalents of silver nitrate in in situ (in situ ) may be transformed into the corresponding (1,2-cyclohexanediamine) diaquaplatinum (II) complex. Subsequently, [Pt (1,2-cyclohexanediamine) (H ₂ O) ₂ ] of the Diaquaa species is transformed into cis -dichloro [Pt (1,2-cyclohexanediamine) Cl ₂ ] complex by adding potassium chloride. Can be.

It will be appreciated that other suitable reaction conditions may be used. In a non-limiting example, the starting material 1,2-cyclohexanediamine-platinum (II) complex is reacted with excess pyrophosphate and the temperature is from about 35 ° C. to about 45 ° C. or any desired narrow between 35 ° C. and 45 ° C. It can be a range. Excellent results were obtained at 40 ° C. In some examples, the reaction may be any desired narrow range between about 13 hours and about 16 hours or between about 13 hours and about 16 hours. Excellent results were obtained at a reaction time of 15 hours. In some examples, the pH may be about 6 to about 7, about 7 to about 8 and about 8 to about 9. Excellent results were obtained at about pH 8.

The aqueous reaction mixture can be concentrated such that no precipitate of pyrophosphate is formed. The aqueous reaction mixture can be concentrated in any suitable manner. For example, the aqueous reaction mixture can be concentrated by rotary evaporation.

As a result, the pH of the reaction mixture can be quickly lowered to less than pH 2 by the addition of a suitable acid. In some instances, nitric acid can be used to lower the pH. In some embodiments, the pH is in the range of about 1 to about 2. Excellent results were obtained at pH 1.

In some examples, the reaction mixture may be cooled to a temperature between 5 ° C. and room temperature (25 ° C. ± 2 ° C.) after concentrating the reaction mixture. In another example, the method also includes lowering the pH of the reaction mixture and then cooling the reaction mixture to a temperature between 5 ° C. and room temperature.

In order to prepare the platinum (IV) complexes according to formulas (II) and (V), additional steps are required for attaching the hydroxyl ligand. Thus, in addition to the steps described above, the reaction mixture is maintained at a pH of about 7 to about 9 at a temperature of about 30 ° C. to about 60 ° C. for a period of about 12 hours to about 18 hours, and then hydrogen peroxide, and Optionally, a reagent selected from the group consisting of acetate salts, butyrate salts, and salts of alpha-hydroxy acids can be added. Any reagent that may be added together with hydrogen peroxide prior to concentration of the reaction mixture may be selected from sodium salts of sodium acetate, sodium butyrate, amines, and alpha-hydroxy acids. In another example, any reagent added with hydrogen peroxide prior to concentration of the reaction mixture may be potassium acetate, potassium butyrate, any monodentate amine such as ammonia, isopropyl amine, etc., and alpha-hydroxy acids. It can be selected from the potassium salt of.

Composition

In some of the various embodiments, (a) provided phosphaplatin complexes; (b) a pharmaceutically acceptable salt of (a); And (c) at least one of the pharmaceutically acceptable solvates of (a). The composition further comprises one or more pharmaceutically acceptable ingredients selected from carriers, diluents, adjuvants and vehicles, generally referring to inert and nontoxic solid or liquid fillers, diluents or encapsulating materials which are nonreactive with phosphaplatin. It may include. Additives of these types are well known in the art and are further described below in connection with methods of treatment. According to various embodiments, provided compositions comprise one or more of the following:

(i) enantiomerically pure (1 R , 2 R ) -pyrodak-2 or a pharmaceutically acceptable salt or solvate thereof:

(I)

;

;

(ii) the enantiomerically pure (1 S , 2 S ) -pyrodak-2 or a pharmaceutically acceptable salt or solvate thereof:

&Lt; RTI ID = 0.0 &

;

;

(iii) an enantiomeric excess of (1 R , 2 R ) -pyrodak-2 of formula (I) or (1 S , 2 S ) -pyrodak-2 of formula (II) or a pharmaceutically acceptable salt or solvent thereof Enantiomerically rich pyridac-2 with cargo:

(I)

&Lt; RTI ID = 0.0 &

;

;

(iv) cis -pyrodak-2 or a pharmaceutically acceptable salt or solvate thereof:

[Formula III]

;

;

(v) enantiomerically pure (1 R , 2 R ) -pyrodak-4 or a pharmaceutically acceptable salt or solvate thereof:

(IV)

;

;

(vi) Enantiomerically pure (1 S , 2 S ) -pyrodak-4 or a pharmaceutically acceptable salt or solvate thereof:

(V)

;

;

(vii) an enantiomeric excess of (1 R , 2 R ) -pyrodak-4 of Formula IV or (1 S , 2 S ) -pyrodak-4 of Formula V or a pharmaceutically acceptable salt or solvent thereof Enantiomerically rich pyrodark-4 with cargo:

(IV)

(V)

; 및

; And

(viii) cis -pyrodak-4 or a pharmaceutically acceptable salt or solvate thereof:

(VI)

.

.

In some embodiments, provided compositions comprise a complex of formula (or a pharmaceutically acceptable salt or solvate thereof) according to any one of formulas (I)-(VI) VI. In some embodiments, provided compositions are multi-complex mixtures of two or more complexes (or pharmaceutically acceptable salts or solvates thereof) of the formula according to any one of Formulas I-VI. The multicomplex mixture may comprise one, two, three, four, five or six of the compounds according to formulas (I) to (VI), provided that the multicomplex mixture is (1 R , 2 R ) -fatigue Pure racemic mixture of dark-2 and ( 1S , 2S ) -pyrodak-2 or pure la of ( 1R , 2R ) -pyrodak-4 and ( 1S , 2S ) -pyrodak-4 It is not a semi mixture.

How to be considered

In further embodiments, the complexes, compositions or both described above may be used alone or in combination with other pharmaceutically acceptable ingredients in a method of treating a proliferative disease or disorder (collectively “disease”). Provided methods include administering to a subject in need thereof a therapeutically effective amount of the complex or composition described above. The subject can be an animal, such as a mammal, including a human. Proliferative diseases that are considered treatable in humans include ovarian cancer, testicular cancer, small cell lung cancer, non-small cell lung cancer and head and neck cancer, skin cancer, pancreatic cancer, breast cancer, colon cancer, glioblastoma cancer. In some embodiments, the complexes and / or compositions can be used in concurrent therapy involving concurrent or sequential treatment with known platinum-metal drugs such as cisplatin, carboplatin and / or oxaliplatin. In addition, the complexes and / or compositions may be used to treat cancers resistant to treatment by one or more of cisplatin, carboplatin, oxaliplatin and / or antimitotic agents such as taxanes, nucleoside homologs, eg For example in combination with gemcitabine, anthracycline antibiotics such as doxorubicin, or targeted therapeutics such as monoclonal antibodies.

As described herein, complexes of formulas (I)-(VI) have been found to be as effective or more effective as cisplatin and carboplatin, and thus cancer for patients who previously lacked effective alternatives to cisplatin and carboplatin treatment. Provide a method of treatment. However, the patient does not need to be previously treated with cisplatin or carboplatin for treatment with the provided complexes, compositions and methods described herein. Administration of the therapeutic agent may be performed by a medical person in a hospital or other medical facility.

Complexes of Formulas (I)-(VI) may be administered and administered according to appropriate medical practice, taking into account the clinical condition, site and method of administration, schedule of administration, patient age, sex, weight, and other factors known to the physician. Thus, for the purposes of the present application, a pharmaceutical “therapeutically effective amount” is determined by these considerations known in the art. The amount should be effective to achieve improvement, including but not limited to improved survival or more rapid recovery, or improvement or elimination of symptoms and other indicators selected by appropriate measures by those skilled in the art. Complexes of the invention can be administered to animals, including mammals and humans, alone or in compositions. In addition, the complexes of Formulas I-VI may be administered over a very wide range of treatments. As one illustrative example, one or more provided complexes may comprise 5 mg / kg to 50 mg / kg; Alternatively 10 mg / kg to 50 mg / kg; Alternatively 20 mg / kg to 50 mg / kg; Alternatively 30 mg / kg to 50 mg / kg; Alternatively 40 mg / kg to 50 mg / kg; Alternatively, it may be administered at one or more doses of 45 mg / kg to 50 mg / kg. Of course, those skilled in the art will appreciate that the therapeutic dose may vary by the subject being administered, the composition being administered and the subject receiving the complex or composition being administered. Thus, as long as a therapeutic dose of less than 5 mg / kg is considered, a therapeutic dose of more than 50 mg / kg is also contemplated. The dose may be a single dose or multiple doses over a period of days. As an illustrative example, it is contemplated that the complex of Formula I-VI may be administered in one, two, three, four, five, six or more doses per day or more. It is also contemplated that the complex may be administered continuously over one or more days, for example by pump or drip. As another illustrative example, the complex may be administered for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days or more.

In the method of treatment, the complexes of Formulas I-VI can be administered in a variety of ways. It should be noted that they can be administered as complexes and can be administered alone in aqueous solution or in combination with pharmaceutically acceptable carriers, diluents, adjuvants and vehicles in view of the good solubility advantages of these complexes. . The complex can be administered by oral, subcutaneous, or intrathecal and infusion techniques as well as parenteral, including intravenous, intraarterial, intramuscular, intraperitoneal, tonsil and intranasal administration. Implantation of the complex may also be useful.

When the complexes of formulas (I) to (VI) are administered parenterally, they will generally be formulated in unit doses of injectable forms (eg, solutions, suspensions, emulsions). Pharmaceutical formulations suitable for injection include sterile aqueous solutions or dispersions and sterile powders for reconstitution into sterile injectable solutions or dispersions. The carrier may be a solvent or a dispersion medium comprising, for example, water, ethanol, polyols (such as glycerol, propylene glycol, liquid polyethylene glycols, etc.), suitable mixtures thereof, and vegetable oils.

Suitable fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants. Non-aqueous vehicles such as cottonseed oil, sesame oil, olive oil, soybean oil, corn oil, sunflower oil or peanut oil and esters such as isopropyl myristate can also be used as solvent systems for the composition. In addition, various additives may be added that enhance the solubility, sterilization and isotonicity of the composition, including antimicrobial preservatives, antioxidants, chelating agents and buffers. Prevention of microbial action can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form may be brought about by the use of agents delaying absorption such as aluminum monostearate and gelatin. However, any vehicle, diluent or additive used must be compatible with the phosphaplatin complex.

Sterile injectable solutions can be prepared by incorporating the phosphaplatin complex in the required amount in a suitable solvent with one or more other ingredients, if desired.

Pharmacological formulations comprising phosphaplatin may be administered to the patient in any suitable carrier, for example an injectable formulation comprising various vehicles, adjuvants, excipients and diluents; Phosphoplatin complexes are in the form of sustained release subcutaneous implants or targeted delivery systems such as monoclonal antibodies, vectored delivery, iontophoretic, polymeric mattresses, liposomes and microspheres. Can be administered parenterally to a patient. Many other implants, delivery systems, and modules are well known to those skilled in the art.

Example

The described embodiments will be better understood with reference to the following examples, which are provided by way of illustration and those skilled in the art will recognize that they are not limiting.

Example  One

(One R ,2 R ) - Fatigue dark -2 Synthesis of Formula I

As starting material for forming the platinum (II) complex, K ₂ PtI ₄ , or more preferably K ₂ PtCl ₄ is reacted with (1 R , 2 R )-(-)-1,2-cyclohexanediamine, respectively to the cis-di-iodo-or cis-dichloro-a (trans - (1 R, 2 R) - (- -) 1,2- cyclohexane-diamine) platinum (II) was formed. Then cis -diiodo-((1 R , 2 R )-(-)-1,2-cyclohexanediamine) platinum (II) or preferably cis -dichloro-((1 R , 2 R )- (-)-1,2-cyclohexanediamine) platinum (II) was dissolved in distilled water (pH 8) with sodium pyrophosphate decahydrate (pH 8) and the resulting mixture was incubated at 40 ° C for 15 hours. After the incubation period, the solution was concentrated by rotary evaporation and filtered to remove any unreacted starting material. The product precipitated by quickly lowering the pH to about 1 by addition of 1N nitric acid. Cooling at about 0 ° C. completes the precipitation and the product is separated by vacuum filtration and washed with cold water and acetone. This synthesis yielded enantiomerically pure (1 R , 2 R ) -pyrodak-2.

Example  2

(One S ,2 S ) - Fatigue dark -2 II ] Synthesis of

Cis -dioyo- or Cis-dichloro- (trans - (1 R, 2 R) - (-) - 1,2- cyclohexane-diamine) system, respectively in place of the platinum (II) - di-iodo-or Cis-Dichloro-the method described in (trans-1,2-cyclohexane diamine, (1 S, 2 S) - - (+)) , except that a platinum (II) as a starting material, in Example 1, Enantiomerically pure (1 S , 2 S ) -1,2-cyclohexanediamine (pyrophosphato) platinum (II) was prepared in a similar manner. This synthesis yielded enantiomerically pure (1 S , 2 S ) -pyrodak-2.

Example  3

(One R ,2 R ) - Fatigue dark -4 IV ] Synthesis of

Starting material from Synthesis Example 1, ie cis -dioodo- or Cis-Dichloro-dissolved (trans - (1 R, 2 R) - (- -) 1,2- cyclohexane-diamine) platinum (II) and pyrophosphate 10-hydrate to distilled water (pH 8), and the resulting mixture Incubate at 40 ° C. for 15 hours. After the incubation period, an aliquot of 30% H ₂ O ₂ was added to the reaction mixture and the reaction mixture was reacted for an additional 3 hours. The solution was then concentrated by rotary evaporation and filtered to remove any unreacted starting material. The product precipitated by the rapid decrease of pH to about 1.0 by addition of 1N nitric acid. Cooling at about 0 ° C. completes the precipitation and the product is separated by vacuum filtration and washed with cold water and acetone. This synthesis yielded enantiomerically pure (1 R , 2 R ) -pyrodak-4.

Example  4

(One S ,2 S ) - Fatigue dark -4 Synthesis of Formula V

Cis-di-iodo-or cis-dichloro- (trans - (1 R, 2 R) - (-) - 1,2- cyclohexane-diamine) platinum (II), respectively, instead of the starting material in Example 2 resulting in, i.e., cis-and is, except for the use of the platinum (II) as the starting material di-iodo-or cis-dichloro- (1,2-cyclohexane-diamine trans - (1 S, 2 S) - - (+)) synthesis example 3 the procedure as pure enantiomerically in a similar manner to (1 S, 2 S) -1,2- cyclohexane diamine described -trans- the dihydric rokso (POS fatigue pato) platinum (IV) was prepared . This synthesis yielded enantiomerically pure (1 S , 2 S ) -pyrodak-4.

Example   5

Sheath - Fatigue dark -2 III ] Synthesis of

In a 500-mL round bottom flask with a stir bar, sodium pyrophosphate decahydrate (0.400 g) was dissolved in distilled water (250 mL). The pH of the solution was then adjusted to 8.0 using 2M nitric acid. The solution was then placed in a 40 ° C. water bath and stirred with a magnetic bar. To the stirred solution was then added cis - dichloro- ( cis -1,2-cyclohexanediamine) platinum (II) (0.100 g, 0.26 mmol). After the mixture was reacted for 15 hours, the solvent was evaporated in vacuo at 48 ° C. to a volume of 5 mL. The mixture was then passed through filter paper and the solution collected in a 10 mL vial with stir bar. The vial was placed in an ice bath on a stir plate and the pH was adjusted from initial 6.5 to 2.0 using 2N nitric acid with gentle stirring. When the low pH was reached, precipitation developed slowly. After stirring was continued for an additional 5 hours, the suspension was filtered through a medium-porosity fritted-glass filter of porous medium that was kept on ice before use. The solid was then washed with cold water (2 parts 5 mL) and cold acetone (2 parts 5 mL) and the filter was placed in the dryer overnight. This resulted in a pale yellow powder (0.077 g, 0.16 mmol, 60% yield).

Example  6

Sheath - Fatigue dark -4 VI ] Synthesis of

In a 500-mL round bottom flask with a stir bar, sodium pyrophosphate decahydrate (0.400 g) was dissolved in distilled water (250 mL). The pH of the solution was then adjusted to 8.0 using 2M nitric acid. The solution was then placed in a 40 ° C. water bath and stirred with a magnetic bar. To the stirred solution was added cis - dichloro- ( cis -1,2-cyclohexanediamine) platinum (II) (0.100 g, 0.26 mmol). The mixture was reacted for 15 hours, 3 mL of 30% (w / w) H ₂ O ₂ was added and an additional 3 hours reaction time was given. The solvent was then evaporated in vacuo at 48 ° C. to a volume of 5 mL. The mixture was then passed through filter paper and the solution collected in a 10-mL vial with stir bar. The vial was placed in an ice bath on a stir plate and gently stirred with pH adjusted from initial 6.5 to 2.5 using 2N nitric acid. Immediately after reaching a low pH, precipitation slowly developed. Stirring was continued for an additional 5 hours and then the suspension was filtered through a micro-glass filter of porous medium that was kept on ice before use. The solid was then washed with cold water (5 mL of 2 parts) and cold acetone (5 mL of 2 parts) and the filter was placed in the dryer overnight. This resulted in a white powder (0.120 g, 0.23 mmol, 88% yield).

Example  7

Phosphaplatin  Characteristic

All phosphopalatin synthesized according to the above synthesis example exhibits solubility in excess of 40 mM / L in aqueous solution of neutral pH in PBS and bicarbonate buffer. (1 R , 2 R ) -Pyrodark-2 and (1 R , 2 R ) -Pyridak-4 show significant stability, especially in aqueous solutions at neutral pH. Typically, no degradation is observed within 7 days after dissolving the phosphaplatin compound in water and observing with ³¹ P-NMR spectroscopy.

Compounds prepared according to the synthetic examples were characterized by confirming the isomeric arrangement by circular dichroism (CD) spectroscopy and by composition by ³¹ P-NMR and mass spectrometer. Additional CD spectra are described in US Patent No. 7,700,649] was performed on a trans- (±) -pyrodak-2 racemic mixture prepared according to the method described in the following.

CD spectra were recorded in phosphate buffer (50 mM) at pH 6.8. The (1 R , 2 R )-and (1 S , 2 S ) -forms of both pyridak-2 and pyridak-4 showed optical activity that could contribute to chirality, but the racemic mixture ( trans- (±) -Pyrodark-2) and cis - isomers of both pyridak-2 and pyridak-4 did not show any CD peaks.

The CD spectrum is shown in FIG. 1, where the numbers in parentheses refer to compounds of the formula corresponding to the numbers in parentheses. The concentrations of the compounds analyzed were: (I), 2.7 mM; (II), 2.5 mM; (III), 1.3 mM; (IV), 3.5 mM; (V), 2.9 mM; (VI), 2.7 mM; And trans - (±) - Fatigue was dark -2, 3.5mM.

Example  8

In vitro Efficacy and Cell Viability Assay ( Clonogenic  black)

To determine the relative activity of phosphaplatin of Formulas (I)-(VI), each stereoisomer was tested via an in vitro clonality assay using human ovarian cancer cells, human colon cancer cells, and human head and neck cancer cells. Human ovarian cancer cells A2780 and A2780 / C30 (cross resistance to 30 μM cisplatin and 100 μM carboplatin) were obtained from Dr. Thomas Hamilton (Dr. Thomas Hamilton, Fox Chase Cancer Center, Philadelphia, PA). Cells were cultured monolayer using RPMI 1640 supplemented with 10% fetal bovine serum, 2 mM glutamine, 0.25 units / mL insulin and penicillin / streptomycin (100 units / mL) in a 37 ° C. incubator with 5% CO ₂ gas continuously. It was. The exponential cell growth was maintained by subculturing the cells with 0.0625% trypsin in HBSS.

Half-maximal inhibitory concentration (IC ₅₀ ) values were measured using the Clonogenic Assay or CyQUANT® Cell Proliferation Assay. In a clonogenic assay, for example, cells were attached by plating 500-700 A2780 cells from a single cell suspension into 60 mm Petri plates 24 hours prior to treatment with the platinum compound. Decant the medium on the day of treatment with the platinum compound described in the Synthesis Example above, replace with a suitable concentration of phosphaplatin compound (50 nM to 75 μM) at three different time points and replace the treated cells with 37 for 24 hours. It was placed back in the incubator. Triple plates were set up for each platinum compound concentration. After 24 hours of treatment, the medium containing the platinum compound was decanted and replaced with fresh medium. These plates were placed in a 37 ° C. incubator for 7 days for colony formation.

In CyQUANT cell proliferation assays, IC ₅₀ values were determined by measuring DNA content using CyQUANT® Cell Proliferation Assay Kit (Invitrogen), which includes a green-fluorescent dye that shows strong fluorescence intensity when cellular DNA binds. In these experiments, the desired number of cells were exposed to phosphaplatin at different concentrations for 72 hours prior to measuring DNA content. Since the DNA content is proportional to the number of viable cells, this assay provides a quantitative measure of proliferating cells. This technique is described in Jones et al., "Sensitive determination of cell number using the CyQUANT cell proliferation assay," J. Immunol . Methods , vol. 254, pp. 85-98 (2001).

IC ₅₀ data from Clonogenic assays and / or CyQUANT cell proliferation assays are summarized in Tables 1-5. In Tables 1 to 5, each reported value is estimated to have an error of ± 15% or less of the reported value, except where an actual error value is provided; Unless otherwise specified, data were obtained from clonality assays.

In addition, these isomeric compounds were tested by exposing these isomeric compounds to the following cell lines: UMSCC10b, Panc-1, UMSCC15s, A2780 / C30 and HCC1806 for 144 hours. Isomer (1 R , 2 R ) -pyrodak-2 showed surprisingly good activity compared to racemic trans- (±) -pyrodak-2. For example, (1 R , 2 R ) -pyrodak-2 is 1.7 in the pancreatic cell line Panc-1, compared to an IC ₅₀ value of 18.5 (μM) for racemic trans- (±) -pyrodak-2. an IC ₅₀ value of (μM) was shown; (1 R, 2 R) - fatigue dark-2 in head and neck cancer cell lines UMSCC10b, rac-trans - (±) - fatigue compared to the IC ₅₀ value of 8.9 (μM) for the dark -2, 0.3 (μM) of the IC ₅₀ value; (1 R, 2 R) - fatigue dark-2 in breast cancer cell lines HCC1806, rac-trans - (±) - fatigue compared to the IC ₅₀ value of greater than 30 (μM) for the dark -2, 9.7 (μM) of the IC ₅₀ values were shown.

In clones immunogenic black trans-isomers of (1 R, 2 R) - fatigue dark-2 and (1 R, 2 R) - Dark fatigue -4 isomers racemic mixture trans- (±) - and trans -2 fatigue dark It showed much better activity than-(±) -pyrrodac-4. For example, trans- (±) -pyrodak-4 exhibited an IC ₅₀ value of 180 ± 15 μM when exposed to human ovarian cancer cells (A2780) for 1 hour, while under the same experimental conditions (Table 1) ( For 1 R , 2 R ) -Pyridak-4, an IC ₅₀ value of 40 ± 10 μM was shown for the same cell line. (1 R, 2 R) on a variety of human cancer cell lines - which showed a prolonged exposure to even lower IC ₅₀ values of the isomers, which indicates the potential as an effective anti-cancer drugs of these compounds. For example, (1 R, 2 R) - fatigue dark-2 is the IC ₅₀ values of 2 μM for the IC ₅₀ values and tolerance of human head and neck cancer of 500nM (0.5μM) on human ovarian cells from clone immunogenic black Had

When A2780 cells are exposed to enantiomerically pure (1 R , 2 R ) -pyrodak-2 and (1 S , 2 S ) -pyrodak-2 compounds for at least 24 hours, both of these compounds are within experimental error. Shows the same activity. However, when cells were exposed for a shorter time, for example 1 hour, different activities were observed between the (1 R , 2 R ) -form and the (1 S , 2 S ) -form. (1 R, 2 R) - form (1 S, 2 S) at a quite short time exposure - shows excellent activity as compared to the isomer, which is due to the cells (1 R, 2 R) - shows a faster absorption of the isomers . (1 R, 2 R) - form or (1 S, 2 S) - compared to the form of the racemic form a high IC ₅₀ value than that of the two types of self-absorbed by the cell to the possibly reduced speed - indicate the federation Can be.

New ( de novo ) For Cisplatin-Resistant Human Ovarian Cancer (OVCAR-10), (1 R , 2 R ) -Pyridak-2 showed better in vitro efficacy than (1 S , 2 S ) -Pyridak-2. It is notable that had exhibited a fatigue dark -2, human ovarian cancer cell line OVCAR-10 (Table 2 and Table 5) and OVCAR-8 (Table 5) markedly superior activity than cisplatin in - (1 R, 2 R) .

Cis -isomers showed superior activity against human ovarian cancer (A2780) compared to other compounds. For example, cis-fatigue dark-2 and cis-fatigue dark-4 corresponding (1 R, 2 R), which after exposure to 24 hours A2780 cell-fatigue dark -2 or (1 R, 2 R) - fatigue, respectively, compared to the IC ₅₀ values of 18μM and 1.0μM against dark -4 shape represents the IC ₅₀ values of 300nM and 4μM, respectively. Note that cisplatin and carboplatin showed much higher IC ₅₀ values, ie, greater than 5.0 μM and 60 μM, respectively, under the same conditions.

Without being bound or limited by theory, the IC ₅₀ values of the chemical principle that expected racemic form (1 R, 2 R) - isomer and the (1 S, 2 S) - isomers arithmetic for each of the IC ₅₀ value means and Teach that they should be the same. As shown in the table, the racemic form of the IC ₅₀ values (for example, IC ₅₀ value of dark -2 fatigue A2780) is (1 R, 2 R), however, - enantiomer and the (1 S, 2 S) - much higher than the expected value, based on a 50/50 mixture of enantiomers. These data suggest that racemic morphology is less potent than expected based on the enantiomer distribution for unknown causes. One valid explanation is that racemic forms may be self-associated and not effectively absorbed by the cells.

In a further test, cells were treated with platinum compound continuously for 7 days. Colonies were fixed and stained using 2% crystal violet in 4% formaldehyde. Colonies containing more than 50 cells were counted. The number of colonies counted from triplicate plates was averaged and this number was divided by the number of smeared cells to obtain fractional values of cell-forming colonies. The fractional values of the cell forming colonies were then corrected for smear efficiency by dividing the fractional values of these cell forming colonies by the number of cell forming colonies in the plate not treated with the platinum compound. Data for colony formation using A2780 is shown in FIG. 2. Both (1 R , 2 R ) -pyrodak-2 and (1 S , 2 S ) -pyrodak-2 enantiomers are 1.0 ± 0.1 μM (1 R , 2 R ) and 1.1 ± 0.1 μM (1 S , 2 S ) showed almost the same IC ₅₀ value, whereas the racemic mixture showed a much higher IC ₅₀ value of 2.4 ± 0.2 μM, indicating the low potency of the racemic mixture. (1 R, 2 R) - Dark fatigue -2 (RD2), (1 R , 2 R) - Dark fatigue -4 (RD4), trans- (±) - fatigue dark -2 (T-D2) and trans- Similar data for OVCAR-10, comparing (±) -Pyrodark-4 (T-D4), are shown in FIG. 3.

Clinical studies are, with reference to the same 1,2-diamino cyclohexane carrier ligand in oxaliplatin, as present in the phosphine papeul Latin described herein, the oxaliplatin (1 R, 2 R) - enantiomer of oxaliplatin other meoga It has been shown to show better efficacy than stereoisomers. In contrast to oxaliplatin, all three pyrophosphato isomers of both pyridak-2 and pyridak-4 (ie, (1 R , 2 R )-, (1 S , 2 S ) -and cis ) It is very active against cancer. However, this does not appear to be a universal trend. For example, as described above, (1 R , 2 R ) -pyrodak-2 and (1 S , 2 S ) -pyrodak-2 are those cells that are exposed to human A2780 cancer cells for 24 hours. Showed almost the same IC ₅₀ value, whereas (1 R , 2 R ) -pyrodak-2 showed better in vitro efficacy for novel cisplatin-resistant human ovarian cancer (OVCAR 10). On the other hand, (1 S, 2 S) - Dark -4 fatigue are all human ovarian cancer cells (1 R, 2 R) in - showed excellent activity than the fatigue dark -4. Cis -isomers of pyridak-2 and pyridak-4 showed better activity against human ovarian cancer (A2780) than trans -isomers. Thus, collectively, these data indicate that some specific cancers can be treated by certain isomers at lower doses compared to racemic forms by avoiding toxicity from other isomers present in racemic forms.

Example  9

On immunofluorescence  by Morocco  Overexpression monitoring

Human ovarian cancer cells A2780 and A2780 /, obtained from Dr. Thomas Hamilton (Dr. Thomas Hamilton, Fox Chase Cancer Center, Philadelphia, PA) in 2.5 mL of medium in 6-well plates with loose, pretreated cover slip. C30 (cross resistant to 30 μM cisplatin and 100 μM carboplatin) was planted. Cells were supplemented with 10% fetal bovine serum, 2 mM glutamine, 0.25 unit / mL insulin and penicillin / streptomycin (100 unit / mL) (Fisher Scientific, Pittsburg, PA) in a 37 ° C. incubator with 5% CO ₂ gas continuously. Cultured as a monolayer using RPMI 1640.

Cells with 70% confluency were treated with one of the phosphaplatin compounds from the synthesis example for 1 hour. The plates were then carefully washed twice with ice-cooled phosphate-buffered saline (PBS), replaced with regular medium and incubated with 37% CO ₂ for another 1 hour. The cover slip was washed twice with PBS and the cells were treated with freshly prepared 1% formaldehyde and then incubated at room temperature for 5-7 minutes. Fixed cells were washed three times with PBS, blocked with 2% FBS / PBS at 4 ° C. for 30 minutes, and then cells were washed three times with PBS for 5 minutes for each wash.

Removing the respective cover slip, and 1 of 5% BSA / 1% milk / PBS on Parafilm ^® surfaces at room temperature for 1 h: a turned FAS primary antibody (Cell Signaling Technology Inc., Danvers, MA) 100μL of diluted 100. The cover slips were then transferred back to clean 6-well plates for each subsequent wash, ie three washes, with PBS for 5 minutes each. The cover slip was passed back to another clean Parafilm ^® surface and incubated with 100 μL of secondary FITC-antibody at 1: 500 dilution in 5% BSA / 1% milk / PBS for 1 hour at room temperature. The cover slips were then washed three times for 5 minutes each with PBS. The moist cover slip is then mounted on the microscope slide with Ultracruz ^® mounting medium with DAPI (4 ', 6-diimidino-2-phenylindole) (Santa Cruz Biotechnology, Santa Cruz, CA) to identify the cell nuclei. It was. Microscopy was performed with fluorescence microscopy at 10 ×, 40 ×, and 100 × magnifications.

Example  10

For protein expression Western Blot / Immunity decoration

After 12% SDS-PAGE electrophoresis, proteins were transferred to PVDF membrane using 100V for 1.5 hours at 4 ° C. The membrane was washed with 0.05% TWEEN-20 and Tris-buffered saline (TBST) solution and blocked with 5% skim dry oil and 1% BSA. Membranes were incubated overnight at 4 ° C. with a primary antibody 1: 25,000 dilution of the protein of interest. The membrane was washed with 0.05% TBST and then incubated with the corresponding HRP-conjugated antibody (1: 40,000 dilution). Proteins were visualized using an ECL-Advanced Chemiluminescent System (ECL-Advance chemiluminescent system, Amersham-GE Healthcare Biosciences, Pittsburg, PA).

Phosphoplatin activates a number of apoptosis and tumor suppressor genes such as FAS, PTEN, PUMA, BAX and the like. Experiments with western blot confirm high levels of protein expression transcribed by these genes. For example, trans- (±) -pyrodark-4 treated mice receive FAS (up to 25 times) and BAX (up to 4 times) upon exposure to trans- (±) -pyrodark-4 for 1 to 12 hours. , Upregulation of PUMA (up to 5 fold) and down regulation of VEGFR (up to 50%). Likewise, BCL2 was downregulated by 70% by RR-Pyrodark-2 and RR-Pyrodark-4. In addition, FAS, FADD and platinum compounds were co-localized in lipid rafts. This activation is also associated with increased expression of sphingomyelinase (Smase), as evidenced by increased protein expression of SMase.

Smase hydrolyzes sphingomyelin into ceramide and phosphorylcholine. In a typical experiment, cancer cells (1 × 10 ⁶ ) were exposed to (1R, 2R) -pyrodak-2 or (1R, 2R) -pyrodak-4 (10 μM) at different time intervals (5 minutes to 2 hours). . Cells were centrifuged and cell lysates collected and washed with cold PBS. Smase activity was monitored using an Amplex red reagent kit (Invitrogen). This assay is performed based on the recommended protocol included in the kit. Typically 11 μL of cell lysate was suspended in 50 mM sodium citrate buffer (pH 5.0) on a 96-well plate and sphingomyelin (5.0 mM) was added to each well. Samples were incubated at 37 ° C. for 1 hour. After incubation, Amplex Red (2 units / mL) horseradish peroxidase, 0.2 units / mL choline oxidase, and 8 units / mL alkaline phosphatase (pH 8.0) were added to each well. The sample was then incubated at 37 ° C. for 30 minutes. Fluorescence intensity was measured at 590 nm using an excitation wavelength of 530-560 m. Fluorescence values from wells containing control samples (untreated) were subtracted from the measurements of each sample.

Example  11

Geology In the raft  Measurement of platinum

Quantitative measurement of platinum content on a Graphite Furnace Atomic Absorption Spectrometer (Perkin Elmer AA-600) from calibration curves established using platinum standards in 0.1% HNO ₃ (Perkin Elmer, Waltham, Mass.) It was. Treated phosphaplatin cells were washed with 1 mL of ice-cooled PBS, stopped protease inhibitor, PI (Pierce, Rockford, IL) four times, and centrifuged at 1000 × g at 4 ° C. between each wash to pellet the pellets. Collected. Cell pellets were added to 250 μL of PBS / PI and protein content was measured using the micro BCA method (Pierce, Rockford, IL). Standard curves were plotted using bovine serum albumin (BSA) prepared at different concentrations. Quantified protein samples were digested in concentrated HNO ₃ for 4 hours and then treated with 30% H ₂ O ₂ for 1 hour before analysis.

Example  12

SCID  Toxicity Studies in Mice

Female SCID mice of 4 to 5 weeks of age (cb-17 / LCR-Prkdc (SCID) / Crl, Strain Code 236 ^r , Charles River Labs, Wilmington, Mass.) Were acclimated for 1 week prior to the start of toxicity testing. Mice were injected intraperitoneally (i / p) of one of the phosphaplatin compounds described in the synthetic examples above in sterile filtered PBS via a sterile 26-gauge needle and syringe. Injections were made at doses of 5 mg / kg to 60 mg / kg and once daily, once every three days and once every five days.

To assess the toxicity of the phosphaplatin compound, side effects (ie, greater than 20% weight loss and / or change in food consumption, deviations from normal behavior, other health problems, or death) were recorded daily. The incidence and severity of adverse events in mice injected with the phosphaplatin compound were compared against the same in mice in the control group. Controls were provided with injections consisting of mock injections (controls for stress response) or PBS (vehicle).

Two groups of mice were treated with a commercially available platinum compound at a dose based on previously published data (ie 12 mg / kg cisplatin and 60 mg / kg carboplatin) for comparison with the phosphaplatin compound. . At the end of the study, all mice were anesthetized with 358 mg / kg avertin and blood was collected by cardiac puncture using a 26-gauge needle and tuberculin syringe. Thus, mice were euthanized by exsanguination under anesthesia. After euthanasia, organs including liver, pancreas, heart, lung, ovary and kidney were harvested, stored in 10% formalin and paraffin blocked for histopathology examination. Changes in tissue characteristics were examined by an experienced pathologist.

SCID mice with ovarian tumors were treated with various doses of trans- (±) -pyrodak-2 and trans- (±) -pyrodak-4 at up to 40 mg / kg and tumor growth was up to 6 weeks or less in these animals The tumor was monitored until the tumor size increased so that they were sacrificed. Tumor growth or regression by phosphatlatin-treated mice was then compared to cisplatin-treated mice (7 mg / kg) and carboplatin-treated mice (60 mg / kg). Three doses of the platinum compound were administered every other day when the tumor grew to at least 100 mm ^{3 in} size. The gross log cell killing value and net log cell killing value were then calculated using the following equation:

Total log cell death rate = ((T-C)) / (3.32 Td)

Pure Log cell death rate = (T-C-treatment period) / 3.32 Td

In the above equations, T and C are the median time (days) for tumors to grow to a specific size, and Td is the median time (days) for doubling tumor size in control animals.

The total log cell killing values recorded for the three dose-therapys were found to be greater than 3, and the pure log cell killing values exceeded 2 at the 40 mg / kg dose. In contrast, cisplatin (7 mg / kg) treated mice, although showing tumor regression, died within 7 days. Carboplatin treated mice (60 mg / kg dose) showed much lower log cell killing values (less than 2) compared to trans- (±) -pyrrodac-4. No further carboplatin dose increase was possible because more than 50% of the population died at the 60 mg / kg dose. Based on in vitro data, in particular cis - pyridak-2, cis -pyrodak-4, (1 R , 2 R ) -pyrodak-2 and (1 R , 2 R ) -pyrodak-4 are racemic It is believed to exhibit better efficacy than trans- (±) -pyrrodac-4.

Example  13

For ovarian cancer cell line A2780 In vivo  efficacy

Stable clonal ovarian cancer cell line A2780 was grown in cell culture until reaching 80-90% confluency. Adherent cells were detached using trypsin (GIBCO / BRL, Grand Island, NY). Trypsinized cells were washed thoroughly with PBS (phosphate buffered saline) to remove trypsin. Human ovarian cancer cell line A2780 SCID atrichia handy gyogye, a SHO-Prkdc ^scid Hr ^hr subcutaneous xenograft by effect of the force papeul Latin compound in (SCID Hairless OutBred, SHO-Prkdc ^scid Hr ^h, Charles River Labs, Wilmington, MA) mouse Evaluated. Female SCID mice, 4-5 weeks of age, were acclimated for 1 week prior to the start of the efficacy test.

Reproduce the cancer cells were suspended in PBS, 1x10 in PBS in all mice except the negative control, mouse cells /0.10mL ⁶ to 5x10 ⁶ cells injected subcutaneously using a 26-gauge sterile needle and syringe into /0.10mL, and a function of time Tumors were evaluated as. Mice were examined daily for tumor growth. Tumors were measured using digital calipers. Tumor Volume Equation (W ² X L) / 2, where W is the tumor measurement at the widest point, L is the tumor dimension at the longest point, and the tumor volume of mm ³ is equivalent to the weight of mg.

About two weeks after subcutaneous injection of cancer cells, the tumors reached a distinguishable tumor size (about 100-200 mm ³ ) and the phosphaplatin compound administration or control injection was initiated. Mice were administered phosphaplatin compound intraperitoneally once a day, once every three days and once again every five days. Each group of xenograft mice was treated with phosphaplatin compound and the corresponding set was treated with vehicle / placebo (PBS solution) and mock injection (stress response control). In addition, two groups of mice were treated with doses based on previously published data (ie 12 mg / kg cisplatin and 60 mg / kg carboplatin) using commercially available platinum compounds to compare with phosphaplatin compounds. .

Daily health monitoring of any adverse health effects of these mice was measured by measuring weight loss / increase and food consumption of xenograft SCID mice. If the tumor size exceeded 3000 mm ³ , the measurement was stopped and the study finished. At the end of the study, all mice were anesthetized with 358 mg / kg avertin and blood was collected by cardiac puncture using a 26-gauge needle and tuberculin syringe. Thus, mice were bled under anesthesia and euthanized. After euthanasia, organs including liver, pancreas, heart, lung, ovary and kidney were collected, stored in 10% formalin and paraffin-blocked for histopathology examination. Changes in tissue characteristics were examined by an experienced pathologist.

Efficacy data of (1 R , 2 R ) -pyrodak-2 and (1 R , 2 R ) -pyrodak-4 on human ovarian cancer cell line A2780 in mouse xenograft model (immune impaired NIH III mice) is shown in FIG. 4. It is summarized in During the test, mice were treated every other day (“qodx3”) (once per day, once every three days and once every five days) for three days. In particular, mice were treated with 40 mg / kg of (1 R , 2 R ) -pyrodak-2, 10 mg / kg of (1 R , 2 R ) -pyrodak-4, or 5 mg / kg of cisplatin. The number N in the legend of FIG. 4 represents the test number and the data points shown beyond the N number were derived as the mean. (1 R, 2 R) - fatigue dark-2 and (1 R, 2 R) - fatigue from the first 10 days of data of Figure 4 of the process to the dark -4 shows clearly the tumor regression during the initial stage of the process.

Example  14

For human ovarian cancer and human head and neck cancer In vivo efficacy

Human ovarian cancer cell line (OVCAR-10) is known to be resistant to both cisplatin and carboplatin. The stable human clonal ovarian cancer cell line OVCAR-10 and the head and neck cancer cell line UMSCC10b were grown separately in cell culture until reaching 80-90% confluency. Adherent cells were detached using trypsin (GIBCO / BRL, Grand Island, NY). Trypsinized cells were washed thoroughly with PBS (phosphate buffered saline) to remove trypsin. From OVCAR-10 and UMSCC-10b xenografts the SCID atrichia handy ^{gyogye, SHO-Prkdc scid Hr hr (} Charles River Labs, Wilmington, MA) mice, NIH (NIH III: NIHBNX- F; NIHS-Lystbg Foxn1nu Btkxid, 4 weeks of age The efficacy of the phosphaplatin compound was evaluated by subcutaneous transplantation into female, Taconic (Rensselaer, NY) mice.

Female SCID / NIH mice, 4-5 weeks of age, were acclimated for 1 week prior to the start of the efficacy test. Cancer cells were resuspended in PBS and all mice except negative control mice were injected subcutaneously with 1 × 10 ⁶ cells / 0.10 mL to 5 × 10 ⁶ cells / 0.10 mL in PBS. Tumor size was assessed as a function of time. Mice were examined daily for tumor growth. Tumors were measured using digital calipers. Tumor Volume Equation (W ² x L) / 2, where W is the tumor measurement at the widest point, L is the tumor dimension at the longest point, and the tumor volume in mm ³ is equivalent to the weight of mg.

About two weeks after subcutaneous injection of cancer cells, the tumors reached a distinguishable tumor size (about 100-200 mm ³ ) and were administered intraperitoneally with the desired dose of phosphaplatin compound. This administration was carried out once a day, once every three days and once again every five days.

Each group of xenograft mice was treated with phosphaplatin compound and the corresponding set was treated with vehicle / placebo (PBS solution) and mock injection (stress response control). In addition, two groups of mice were treated with doses based on previously published data (ie 12 mg / kg cisplatin and 60 mg / kg carboplatin) using commercially available platinum compounds to compare with phosphaplatin compounds. . The health side effects of these mice were monitored daily by measuring weight loss / increase and food consumption of xenograft SCID mice / NIH. If the tumor size exceeded 3000 mm ³ , the measurement was stopped and the study finished.

 At the end of the study, mice were anesthetized with 358 mg / kg avertin and blood was collected by cardiac puncture using a 26-gauge needle and tuberculin syringe. Mice were bled under anesthesia and euthanized. After euthanasia, organs including liver, pancreas, heart, lung, ovary and kidney were harvested, stored in 10% formalin and paraffin blocked for histopathology examination. Changes in tissue characteristics were examined by an experienced pathologist.

Human ovarian cancer cells for the OVCAR-10 (1 R, 2 R) - fatigue dark-2 and (1 R, 2 R) - efficacy data of fatigue dark -4 are summarized in Figure 5 as compared to PBS / Bicarb control have. 40 mg / kg body weight was administered when tumors became 100-200 mm ^{3 in} size. Treated mice showed tumor regression during the treatment time. Tumor size measurements over time for qodx3 control (PBS / bicarbonate), 60 mg / kg carboplatin with qodx3 and 40 mg / kg with qdx3 were administered over (1 R , 2 R ) -pyrrodac-4 It is summarized in FIG. The dashed arrows highlight the time (days inserted if necessary) that the mean tumor size reached 1000 mm ³ . Compared to the control, carboplatin showed an increased lifespan (% ILS) of only about 125%, while ( 1R, 2R ) -pyrodak-4 showed about 209%% ILS.

Efficacy data of (1 R , 2 R ) -pyrodak-2 and (1 R , 2 R ) -pyrodak-4 against human head and neck cancer UMSCC10b are summarized in FIG. 7. Doses ranging from 10 mg / kg body weight to 40 mg / kg body weight were administered qodx3 and / or qdx4. These potencies were compared to the racemic form trans- (±) -pyrrodac-4. The dose was administered when tumors reached a size of 100-200 mm ³ .

Example  15

(One R ,2 R ) - Fatigue dark -2 and (1 R ,2 R ) - Fatigue dark -4 max Tolerance  Volume

To determine the maximum tolerated dose, (1 R , 2 R ) -Pyrodark-2, (1 R , 2 R ) -Pyridak-4, trans- (±) -pyrodak-4, trans- (±) Toxicity experiments were performed on pyrodak-4, cisplatin and carboplatin. Four to five week old female ICR (CD-1) mice (Taconic (Rensselaer, NY)) were dosed with increasing doses from 10 mg / kg to 100 mg / kg. Mice weighed 15 g to 24 g. Mice were injected with one of the phosphaplatins every other day in a three-dose regimen. At least 20% total weight loss, poor growth, weight gain failure, other observable health problems, departure from normal behavior or death were considered side effects.

No weight loss was observed at low doses of 10 mg / kg. No death or 20% total weight loss was observed for all phosphaplatin compounds up to the highest dose of 40 mg / kg. These results can be compared with cisplatin, in which 100% of mice died at a dose of 12 mg / kg, and carboplatin, where 33% of deaths were observed at a dose of 60 mg / kg. At high doses of 100 mg / kg, all mice showed greater than 20% weight loss or died within 15 days of administration.

Example  16

real time PCR  Quantitative Gene Expression from Experiment

Quantitative real-time PCR experiments were performed to assess the expression of a few target genes. Human epithelial ovarian cancer cell line (A2780) and cisplatin-resistant cell line (A2780 / C30) were treated with cisplatin, (1 R , 2 R ) -pyrodak-4 and trans- (±) -fatigue for 0, 3, 12 and 24 hours. Incubations were made in RPMI 1640 with or without dark-4. Both treated and untreated cancer cells were maintained in RPMI 1640 medium with 10% fetal bovine serum, 2 μmol L-glutamine, 100 units / mL penicillin-streptomycin and 0.25 units / mL insulin solution at 37 ° C. with 5% CO ₂ . RNA was isolated from both treated and untreated cells. Treated cells are cells harvested after exposure to an IC ₅₀ value concentration of phosphaplatin at different time intervals from 3 to 24 hours.

RNA samples were treated with DNase-free RNase (QIAGEN Sciences) to remove DNA. The cDNA was then synthesized using the High Capacity RNA-to-cDNA kit (Applied Biosystems) according to the manufacturer's instructions. The amount and purity of RNA was measured by UV spectroscopy (NanoDrop 2000 Thermo Scientific, Wilmington, DE). A minimum absorbance ratio indicator of 1.9 (the ratio of absorbance measured at 260 nm to the absorbance measured at 280 nm) was used as an acceptable purity.

Separated RNA samples were stored at −80 ° C. and treated with a minimum freeze-thaw cycle to maintain RNA integrity. Double-time real-time PCR was performed on target genes and endogenous controls (β-actin) in the same reaction wells using the Taqman ^® gene expression assay. Endogenous controls are the criteria used to normalize target mRNA. This chain reaction was initially performed using different cDNA concentrations to determine the optimal concentration of cDNA needed to detect the gene of interest. Target genes were labeled with blue dye (FAM, absorbance: 494 nm, emission: 518 nm), while reference genes were tagged with green dye (VIC, absorbance: 538 nm, emission: 554 nm). cDNA concentration is determined by ABI StepOnePlus ^™, where the threshold cycle (Ct) value for the target gene is used for measurement. It was chosen to be 17-32 most sensitive to fluorescence detection by the instrument.

The threshold cycle number (Ct) was measured at which point the fluorescence of the qPCR reaction just exceeded the threshold fluorescence of the detection system. These Ct values were used to compare with the levels of target genes and endogenous controls. Fold (fold), which is calculated from (ΔCt) ΔCt and ΔΔCt value corresponding to _{0 hours (control)} - - changing (2 Quantitative gene expression, a relational expression ΔCt = (Ct _targets - _see Ct) and ΔΔCt = (Δt) _{of time X} ^{- ΔΔ Ct} ). The fold-expression for the control sample (untreated) was still equal to 1 because the ΔΔCt value was 0 and therefore 2 ⁰ was 1.

This application should not be considered limited to the specific examples and embodiments described herein, but rather should be understood to cover all aspects of the invention. Various modifications, equivalent processes, and various structures and apparatuses to which the present invention is applicable will be readily apparent to those skilled in the art. Those skilled in the art will understand that various changes may be made without departing from the scope of the present invention and are not intended to be limiting as described herein.

Claims (32)

Phosphoplatin complexes selected from the group consisting of:
(i) enantiomeric pure (1 R , 2 R ) -pyrodak-2 or a pharmaceutically acceptable salt or solvate thereof:
(I)

;
(ii) the enantiomerically pure (1 S , 2 S ) -pyrodak-2 or a pharmaceutically acceptable salt or solvate thereof:
[Formula II]

;
(iii) cis -pyrodak-2 or a pharmaceutically acceptable salt or solvate thereof:
(III)

;
(iv) enantiomerically pure (1 R , 2 R ) -pyrodak-4 or a pharmaceutically acceptable salt or solvate thereof:
[Formula IV]

;
(v) enantiomerically pure (1 S , 2 S ) -pyrodak-4 or a pharmaceutically acceptable salt or solvate thereof:
(V)

; And
(vi) cis -pyrodak-4 or a pharmaceutically acceptable salt or solvate thereof:
(VI)

. Phosphoplatin complexes selected from the group consisting of:
(i) an enantiomeric excess of (1 R , 2 R ) -pyrodak-2 of formula I or (1 S , 2 S ) -pyrodak-2 of formula II or a pharmaceutically acceptable thereof Enantioenriched pyridac-2 with salts or solvates which are:
(I)

[Formula II]

; And
(ii) an enantiomeric excess of (1 R , 2 R ) -pyrodak-4 of Formula IV or (1 S , 2 S ) -pyrodak-4 of Formula V or a pharmaceutically acceptable salt or solvent thereof Enantiomerically rich pyrodark-4 with cargo:
[Formula IV]

(V)

. The method of claim 2 wherein the enantiomerically excess (1 R, 2 R) - fatigue consisting of abundant dark fatigue -2 enantiomerically the dark with 0.1% -2% to 99%, force papeul Latin complexes. The method of claim 2 wherein the enantiomerically excess of (1 S, 2 S) - fatigue consisting of abundant dark fatigue -2 enantiomerically the dark with 0.1% -2% to 99%, force papeul Latin complexes. The method of claim 2 wherein the enantiomerically excess (1 R, 2 R) - fatigue consisting of abundant dark fatigue -4 enantiomerically the dark with 0.1% -4% to 99%, force papeul Latin complexes. The method of claim 2 wherein the enantiomerically excess of (1 S, 2 S) - fatigue consisting of abundant dark fatigue -4 enantiomerically the dark with 0.1% -4% to 99%, force papeul Latin complexes. (a)
(i) enantiomerically rich pyridac-2 having an enantiomeric excess of (1 R , 2 R ) -pyrodak-2 of formula I or a pharmaceutically acceptable salt or solvate thereof:
(I)

;
(ii) a thio Ahira meojeok excess of formula II (1 S, 2 S) - Dark -2 fatigue or rich as enantiomers meojeok fatigue with a pharmaceutically acceptable salt or solvate thereof as medicament dark-2:
[Formula II]

;
(iii) cis -pyrodak-2 or a pharmaceutically acceptable salt or solvate thereof:
(III)

;
(iv) enantiomerically rich pyridak-4 with an enantiomeric excess of (1 R , 2 R ) -pyrodak-4 of formula IV or a pharmaceutically acceptable salt or solvate thereof:
[Formula IV]

;
(v) enantiomerically rich pyridak-4 with an enantiomeric excess of (1 S , 2 S ) -pyrodak-4 of formula V or a pharmaceutically acceptable salt or solvate thereof:
(V)

;
(vi) cis -pyrodak-4 or a pharmaceutically acceptable salt or solvate thereof:
(VI)

; And
(vii) at least one phosphatlatin complex selected from the mixture of two or more of the phosphaplatin complexes (i) to (vi); And
(b) a composition for treating a proliferative disease, comprising one or more pharmaceutically acceptable ingredients selected from the group consisting of carriers, diluents, adjuvants, and vehicles. 8. The composition of claim 7, wherein the phosphaplatin complex is an isolated monomeric phosphaplatin complex. The method of claim 7, wherein enantiomerically excess (1 R, 2 R) - blood containing blood rich Dark -2 enantiomerically dark -2 with 10% to 100%, the composition. 10. The composition of claim 9, wherein said enantiomerically rich pyridak-2 consists of (1 R , 2 R ) -pyrodak-2 of formula I and (1 S , 2 S ) -pyrodak-2 of formula II. , Composition:
(I)

[Formula II]

. 8. The composition of claim 7 comprising enantiomerically rich pyridak-2 having an enantiomeric excess of (1 S , 2 S ) -pyrodak-2 from 10% to 100%. 12. The composition of claim 11, wherein said enantiomerically rich pyridak-2 consists of (1 R , 2 R ) -pyrodak-2 of formula I and (1 S , 2 S ) -pyrodak-2 of formula II. , Composition:
(I)

[Formula II]

. 8. The composition of claim 7, comprising enantiomerically enriched pyridak-4 having an enantiomeric excess of (1 R , 2 R ) -pyrodak-4 in a range of 10% to 100%. 14. The composition of claim 13, wherein said enantiomerically rich pyridak-4 consists of (1 R , 2 R ) -pyrodak-4 of formula IV and (1 S , 2 S ) -pyrodak-4 of formula V , Composition:
[Formula IV]

(V)

. 8. The composition of claim 7, comprising enantiomerically enriched pyridak-4 having an enantiomeric excess of (1 S , 2 S ) -pyrodak-4 in a range of 10% to 100%. 16. The composition of claim 15, wherein said enantiomerically rich pyridak-4 consists of (1 R , 2 R ) -pyrodak-4 of formula IV and (1 S , 2 S ) -pyrodak-4 of formula V , Composition:
[Formula IV]

(V)

. The composition of claim 7 comprising cis -pyrodak-2. The composition of claim 7 comprising cis -pyrodak-4. The composition of claim 7, wherein the proliferative disease is cancer. 20. The method according to any one of claims 7 to 19, wherein the proliferative disease is selected from ovarian cancer, testicular cancer, small cell lung cancer, non-small cell lung cancer, head and neck cancer, skin cancer, pancreatic cancer, breast cancer, glioblastoma cancer, and colon cancer. , Composition. The composition of any one of claims 7-19, wherein the proliferative disease is cancer resistant to treatment by one or more of cisplatin, carboplatin and oxaliplatin. (a) (i) enantiomerically rich pyridac-2 having an enantiomeric excess of (1 R , 2 R ) -pyrodak-2 of formula I or a pharmaceutically acceptable salt or solvate thereof:
(I)

;
(ii) a thio Ahira meojeok excess of formula II (1 S, 2 S) - Dark -2 fatigue or rich as enantiomers meojeok fatigue with a pharmaceutically acceptable salt or solvate thereof as medicament dark-2:
[Formula II]

;
(iii) cis -pyrodak-2 or a pharmaceutically acceptable salt or solvate thereof:
(III)

;
(iv) enantiomerically rich pyridak-4 with an enantiomeric excess of (1 R , 2 R ) -pyrodak-4 of formula IV or a pharmaceutically acceptable salt or solvate thereof:
[Formula IV]

;
(v) enantiomerically rich pyridak-4 with an enantiomeric excess of (1 S , 2 S ) -pyrodak-4 of formula V or a pharmaceutically acceptable salt or solvate thereof:
(V)

;
(vi) cis -pyrodak-4 or a pharmaceutically acceptable salt or solvate thereof:
(VI)

; And
(vii) A therapeutically effective amount of a composition comprising at least one phosphaplatin complex selected from a mixture of two or more of the phosphaplatin complexes (i) to (vi) is administered to a subject in need of treatment of a proliferative disease. A method of treating a proliferative disease, comprising administering. The method of claim 22, wherein the composition further comprises one or more pharmaceutically acceptable ingredients selected from (b) carriers, diluents, adjuvants, and vehicles. The method of claim 22 or 23, wherein the composition
(i) enantiomerically pure (1 R , 2 R ) -pyrodak-2;
(ii) enantiomerically pure (1 S , 2 S ) -pyrodak-2;
(iii) enantiomerically pure (1 R , 2 R ) -pyrodak-4;
(iv) enantiomerically pure (1 S , 2 S ) -pyrodak-4;
(v) cis -pyrodak-2; And
(vi) at least one phosphaplatin complex selected from the group consisting of cis -pyrodak-4,
The composition
Does not include both (1 R , 2 R ) -pyrodak-2 and (1 S , 2 S ) -pyrodak-2;
The composition
The method does not include both (1 R , 2 R ) -pyrodak-4 and (1 S , 2 S ) -pyrodak-4. The method of claim 22, wherein the one or more phosphopalatin complexes of the composition are enantiomerically enriched pyridak-2, enantiomerically enriched pyridak-4, or both. Method, 26. The composition of any one of claims 22 to 25, wherein the one or more phosphaplatin complexes of the composition are enantiomerically pure (1 R , 2 R ) -pyrodac-2, enantiomerically pure ( 1 R , 2 R ) -pyrrodac-4 or both. 27. The method of any one of claims 22-26, wherein the composition further comprises cisplatin, carboplatin, oxaliplatin, or a combination thereof. The method of any one of claims 22-27, wherein the proliferative disease is cancer. The method of claim 28, wherein the proliferative disease is selected from ovarian cancer, testicular cancer, small cell lung cancer, non-small cell lung cancer, head and neck cancer, skin cancer, pancreatic cancer, breast cancer, glioblastoma, and colon cancer. The method of claim 28, wherein the proliferative disease is cancer resistant to treatment with one or more of cisplatin, carboplatin, and oxaliplatin. 31. The method of any one of claims 22-30, wherein the subject is a mammal. 31. The method of any one of claims 22-30, wherein the subject is a human.

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